摘要
目的 建立一种检测临床标本中微量梅毒螺旋体 (Tp)的敏感、特异的方法。方法 运用常规及巢式PCR扩增了Tp的DNA多聚酶I基因 (polA)的特异性片段 ,检测了两方法的特异性和敏感性 ;与血清学方法比较 ,评价了两方法在诊断梅毒中的意义。结果 常规及巢式 polAPCR只在扩增Tp时有特异性产物 ,灵敏度分别约为 40个 /10 0 μl及 1个 /10 0 μl。检测 13 1份拟诊为梅毒的血清标本 ,血清学方法与巢式 polAPCR结果无显著性差异 ;巢式和常规 polAPCR结果有显著性差异 ,巢式PCR敏感性高于常规PCR。结论 巢式 polAPCR具有高度敏感、特异、标本处理简便等优点 ,适用于检测血清等标本中微量Tp ,可作为血清学方法的补充用于梅毒诊断。
Objective To develop a sensitive and specific method to detect extremely low numbers of T.pallidum in clinical samples.Methods PCR and nested PCR were performed to amplify specific fragments of DNA polymeraseⅠgene(polA) of T.pallidum. Sensitivity and specificity of the two PCR methods were analyzed.Compared with serologic tests,two PCR methods were evaluated in the diagnosis of syphilis. Results Only specific amplicons could be found in amplifying the T.pallidum polA by two PCR methods.The detection limit was about 40 and 1 organisms per 100 μl reaction by routine PCR and nested PCR,respectively.In the detection of 131 serum specimens from persons with suspected syphilis,no significant differences could be found in the serology and nested PCR.However,there were significant differences between the nested PCR and the routine PCR and the nested polA PCR was much more sensitive than the routine polA PCR.Conclusion The nested polA PCR method is extremely sensitive,specific and simple for detecting low numbers of T.pallidum in clinical serum samples as a significant addition to the serologic tests for syphilis.
出处
《中国皮肤性病学杂志》
CAS
北大核心
2004年第2期74-76,共3页
The Chinese Journal of Dermatovenereology
基金
湖南省教育厅资助重点课题 (0 0 2A0 0 8)
湖南省卫生厅资助课题 (Y0 2 - 0 67)
